Impaired inflammasome activation and bacterial clearance in G6PD deficiency due to defective NOX/p38 MAPK/AP-1 redox signaling

Yen, Wei-Chen; Wu, Yi-Hsuan; Wu, Chih-Ching; Lin, Hsin-Ru; Stern, Arnold; Chen, Shih-Hsiang; Shu, Jwu-Ching; Tsun-Yee Chiu, Daniel

Ficoll-Paque was purchased from GE Healthcare (Little Halfont, Buckinghamshire, UK). Ultrapure LPS was purchased from InvivoGen (California, USA). Anti-IL-1β, anti-p65, anti-phospho-p65, anti-p38, anti-phospho-p38, anti-ERK, anti-phospho-ERK, anti-JNK, antiphospho-JNK, anti-c-Jun, anti-phospho-c-Jun, anti-phospho-c-Fos and anti-c-Fos were all purchased from Cell Signaling Technology (Beverly, MA, USA). Anti-G6PD was purchased from Genesis Biotech (Taiwan). ATP, nigericin, PMA, hydrogen peroxide, DPI, NADPH, and anti-β-Actin were purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-NLRP3 was purchased from AdipoGen Life Sciences (San Diego, CA, USA), and anti-ASC was purchased from Medical & Biological Laboratories (Japan). The anti-caspase-1, anti-pro-IL-1β, anti-mouse and anti-rabbit antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-α-tubulin was purchased from Merck Millipore (Burlington, USA). Anti-lamin B1 was purchased from Proteintech (Illinois, USA). Polyvinylidene difluoride (PVDF) membranes and Immobilon Western Chemiluminescent HRP Substrate (ECL) were purchased from Millipore Corporation (Billerica, MA, USA).